This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Core 2A is developing tools to analyze the Glycoprotein Glycome of cells, including embryonic mouse stem cells and their differentiated products. Fractionation of tryptic glycopeptides using serial lectin affinity chromatography is being developed to classify expression by the types of glycans expressed. Separation of glycopeptides is then further accomplished by liquid chromatography, followed by mass spectrometry to catalog and identify various species. At various stages, N-linked glycans will be produced by N-glycanase treatment. The resulting peptides will be identified by their Asn to Asp conversion using LC/MS analysis. All results will be submitted to the Bioinformatics module (Core project 4) for integration with other data.